This work reports on the fabrication and performance of a simple amperometric immunosensor device to be potentially used for the detection of serum anti-citrullinated peptide antibodies (ACPAs), which are specific for rheumatoid arthritis (RA) autoimmune disease. Sera of RA patients contain antibodies to different citrullinated peptides and proteins such as fibrin or filaggrin. Herein, a chimeric fibrin–filaggrin synthetic peptide (CFFCP1) was used as a recognition element anchored to the surface of a multiwalled carbon nanotube–polystyrene (MWCNT–PS) based electrochemical transducer. The transducer
fabrication process is described in detail together with its successful electrochemical performance in terms of repeatability and reproducibility of the corresponding amperometric response. The resulting
immunosensor approach was initially tested in sera of rabbits previously inoculated with the synthetic peptide and eventually applied to the detection of ACPAs in human sera. A comparative study was carried out using control serum from a blood donor, which demonstrated the selectivity of the immunosensor response and its sensitivity for the detection of anti-CFFCP1 antibodies present in RA patients.